## OTS0523254 

# • N 111111111111111111111111111111111111111111111111111111111111



### One Source. One Search. One Solution. 

# IN VITRO MICROBIOLOGICAL MUTAGENICITY STUDIES OF FOUR CIBA·GEIGY CORPORATION COMPOUNDS (FINAL REPORT) WITH TEST DATA AND COVER LETTER

08 MAR 1990

U.S. Department of Commerce 

### National Technical Information Service

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## More than 600,000 government research information products have been added to the NTIS collection since 1990. All bibliographic entries for those products are searchable on the NTIS Web site at www.ntis.gov.
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## For more information visit our website: 

# www.ntis.gov**/'i~"":\** U.S DEPARTMENT OF COMMERCE



# . 1Jj''

. Technology Administration 

# \ ·r / National Technical Information Service

..._., Springfield, VA 22161

## _C_9PING FORM FOR SRC INDEXING, REVISED 10'115185

IN VITRO MICROBIOLOGICAL MUTAGENICITY STIJDIES OF FOUR CI-...sEISV CORPORATICJt.! COMPOUNDS (FINAL REPORT) WITH TEST DATA AND COVER LETTER

He:XABROMOCVCLODODECANE (3194-55-1!;) Q. 

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# GreafLakes-. Chemical**Corpitratio~·**



### March 8, 1990

Document Processing Center (TS-790) Room L-100, Office of Toxic Substances Environmental Protection Agency 401 M street, sw Washington, DC 20460 Attn: 8 (d) Health & 

### Safety Reporting,, (Notification/Reporting) Dear Sir: 

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# - In response to the TSCA Section 8 {d) rule' published in the December 12, 1989 Federal Register, the enclosed information is being sub- , mitted on the**followin~** 

### substances:

The the 2,4,6-Tribrornophenol 'Tetrabrornophthalic anhydride Hexabromocyclododecane**1,2-Bis(tribrornophenoA~)ethane** 

### Decabromodiphenyl ether Pentabromodiphenyl ether Octabromodiphenyl ether studies on the first ,four compounds following manufacturing facility:

(., 

### Great Lakes Chemical' corporation Route 7, Box 529 El Dorado, AR 71730 cAs Number 118-79-6 632-79-1 3194-55-6 37853-59-1 1163-19-5 32534-81-9**32536-52-0~-** ; 

### :.



### are submitted on behalf of

The studies on the last three cornpou."lds are**su~mitted** 

### on behalf of the fcil:J.owing manufacturing facility: ' •, Great Lakes Chemical Corporation '' Highway 15 South, P.O.Box 1878 · El Dorado, AR 71731-1878 

## 'r;.



### " 

### There are no known additives to these compounds. Pentabromodiphenyl ether consists of hexa-, penta-·, tetra-bromodiphenyl ether. Octabromodi.phenyl ether consists of nona-, octa-., hepta"'-, and hexa- " bromodi.phenyl ether. ·we are not aware of any ongoing studies with any of the listed compounds. If you have any questions concerning this submission, please call me at (317) 497-639l.: 

### ·--. Sincerely,

# J).L-tt"-d/~**/..{<f~** Dennis L. McFadden, Ph. D. Product Safety Coordinator ,. DLM:sb:OJOS90 Enclosures

;.-::. <.:



## ·' 

# .

### II 

### TEST DATA 2,4,6-Tribromophenol Acute toxicity studies in rats and rabbits Primary skin i=itation study in albino rabbits Eye! 

### i=itation study in albino rabbits Acute inhalation toxicity in the albino rat Acute dermal toxicity in male and female albino rabbits 'Acute oral toxicity (LD 50 ) 

### study in albino rats Dermal sensitization study in the albino guinea pig Bioaccumulation study in fat tissue with 2,4,6-tribromophenol 48-Hour dynamic aquatic toxicity study with 2,4,6-tribromo- phenol in Daphnia

**Four-~ay** 

### static aquatic toxicity studies with 2, 4 , 6-tribromo-

.. • phenol in rainbow trout and bluegills. Screening test for microbiological inhibition with 2,4,6- tribromophenol Acute dust inhalation toxicity study with 2,4,6-tribromo- phenol, lot number 3106 in rats · 28-Day subacute dermal toxicity study with 2,4,6-tribromo- phenol in albino rabbits (10/27/77) 28-Day subacute dermal toxicity study wth 2,4,6-tribromo- phenol in albino rabbits (12/12/77) · Pilot teratology study in rats Mutagenicity evaluation of 2,4,6-trib:i:oroophenol, lot number 3287, in the Ames Salmonella/Microsome Plate Test Acute oral toxicity (LD 5 

# J study in rats ' 

### =· The bioaccumulation of 2,4,6-tribromophenol in bluegill sunfish 2,4,6-Tribromophenol biodegradation (9/30/75) 2,4,6-Tribromophenol (TBP) biodegradation (6/1/76) Pharmacokinetics study of 2,4,6-tribromophenol in rats Photolysis of 2,4,6-tribromophenol ·partition coefficient of several flame retardants and industrial chemicals Water solubility of several flame retardants and-\ndustrial chemicals

-- Photolysis of 2,4,6-tribromophenol in aqueous solution



# ,,



# .

**~·.** 

## '

### Tetrabromophthalic Anhydride Acute oral study Acute dermal study A.cute toxicity studies of tetrabromophthalic anhydride Primary skin irri.tation study in albino rabbits Eye irritat:i.'On study in albino rabbits Acute inhalation toxicity in t;he albino rat Dermal sensitization study in the albino guinea pig Acute inhalation toxicity in the albino rat after'pyrolysis (3/12/75) 

## r;?' Acute inhalation toxicity in the albino rat· after pyrolysis (3/26/75) 28-Day dermal toxicity study in rabbits 21-Day inhalation toxicity in rats Human repeated insult patch test with Firemaster PHT4 Mutagenicity_evaluation of 859-74-4 Pilot teratology study in ra't:s ' ' The acute toxicity of Firemaster PHT4 to the water flea,

. Daphnia magna Straus The acute**tox~city** 

### of Fi.,emaster PHT4, lot number 6332-B to the rainbow trout Salmo gairdne:o:i Richardson The acute toxicity of**Fire~aster** 

### PHT4, lot number 6332-B to the bluegill sunfish Lepomis macrohirus Rafinesque Acute oral toxicity study in mice Comparative inhalation toxicity tests**'-~** 

### Evaluation of the inhalation toxicity of combustion products of selected polymers II The. bioaccumulation of tetrabr·omophthalic; anhydride in the bluegill sunfish '' \; Persistence of PHT-4 in soil at 1 and 10 p.gjg

Photolysis of PHT-4 .Effects of PHT-4 on soil microflora.at 1 and 10 p.g/g ''Pr.armacokinetics of PHT-·4 in r-ats

Partition coefficient of several flame retardants and industrial chenicals Water solubility of several flame retardants and industrial chemicals



# ,, (•

::

:;:

D**·~·**



### ! ·: 

### .,

### Hexabromocyclododecane /.Hexabromocyclododecane: 28-day feeding trials with rats and 90-day feeding trials witll rats VIn vitro microbiological mutgenicity studies of four Ciba

### Geigy Corporation compounds Pilot cataractogenic study in chicks cataractogenic study in chicks Mutagenicity evaluation of 421-32B Acute toxicity studies in rabbits ad rats Mutagenicity of two lots of FM-100, lot**~3** 

### and residue of lot 3322 in the absence and presence of metabolic activation ,; .. , ·· The acute toxicity of HBCD lot number 990-17 tq the**bluegi~l** 

### sunfish Lepomis macroh'irus Rafinesque Partition coefficient of dicamba, endrin, VEL 3510 and' several industrial chemicals and flame retardants Water solubility of several industrial chemicals, flame retardants and a herbicide VEL 3510 Hydrolysis of Firemaster 100 HBCD

- biodegradability study

(i

:,: 

## " "

# ,,

,, 1,2-BisCtribromophenoxylethane

l.i 

### -water solubility of several flame retardants and industrial chemicals -Photolysis of Firemaster 680 -Pharmcokinetic study of FM-680 -Rate of hydrolysis studies -Partition coeffi.cient of MC-680, chlorendic anhydride and chlorendic acid -28-Day rat feeding study -Acute oral toxicity study in the albino rat -Acute dermal LD 50 

### (l level) -Acute inhalation toxicity in the albino rat -Acute oral toxicity study 

## in beagle dogs --il.cute toxicity (TL.,; 0 ) 

### study 

## in the bluegill -Acute toxicity (LD 50 ) 

### study in the rainbow trout -Acute heated vap.or inhalation toxicity study in rats -Modified Draize niul tiple

.. insult test in male humans -28-Day dermal toxicity study in rabbits

**~21-Day** 

### inhalation toxicity in rats -Acute inhalation toxicity 

## in the albino rat after pyrolysis (ABS resin/FM-680/Sb 2 

### 0.3) Acute inhalation toxicity 

## in the albknO rat after pyrolysis

**(~BS ;:esin/~b** 

**)** 

### ._14-Day range

**fnd~ng** 

### study Ln rats '28-Day toxicity study in rats 'Biodegradation study with' 14 

### C-tagged MC-680 -Biodegradability study"with 14 

### c-tagged MC-.680 " "Primary skin irr·itation test with mother ·liquor 680 process, code no. 7-29-76-B 

## in albino rabbits -skin sensitization test with mother liquor 680 process in albino guinea pigs """'90-Day subacute oral toxicity study with Firemaster 680 in albino rats (Jan. 6,: 1977) .:• '90-Day subacute oral toxicity study with Firemaster 680 final report 1.";



## in albino rats (Mar. 10, 1977') 'Mutagenicity evaluation of 859-74-5 --Test report (method for testing the of chemical substances in fish in fish) · degree of accumulation boay) (Bioaccuniulatiofi

,,, "'Acute inhalation toxicity study in the albino rat after pyrolysis (ABS resin/Sb 2 0;Vcontaining 680) Pilot teratology study in rats · 'Teratology study in rats ' "Tissue residue accumulation/depletion study with Firemaster 680 in 

### albino rats · ,, 

## ,,

### Decabromodiphenvl ether \Acute toxicity studies in rats and rabbits \Mutagenicity evaluation of componrid 364-5A ; .'-28-Day. toxicity study.

**-in.,rats·..:.·.(-5/-12/-7~).,.'=·'==--=;.-'----"'·**



### '28-Day toxicity study in rats (6/24/76) '28-Day toxicity study in rats (3/2/77)

**~Mutagenicity** 

### evaluation of compound 277-10 Pentabromodiphenyl oxide -r!utagenicity evaluation of compound 345-76A final report, 1 , 

### 'The acute oral toxicity of pentabromodiphenyl ether to rats· \Acute toxicity studies in rats and rabbits \28-Day toxicity study in rats 'The bioaccumulation of compound S-512 by carp "'Jo-oay dietary study in rats with pentabromodipl'i:enyl oxide including recovery periods of 6, 12, and 24 weeks

**~** 

### '90-Day dietary study irrrats with pentabromodiphenyl oxide (DE'-71)
,, 

### Octabromodiphenyl oxide a 

# J The bioaccumulation of compound S-511. by carp ..-1\cute toxicity studies in rats and rabbits v1subacute inh.alation toxicity study in rats

**~Mutagenicity** 

### evaluation of compound

**34S~79A** 

### final report

., Jrn vitro sister chromatid exchange 

## in Chinese hamster• ovary cells with octabromodiphenyl ether (DE-79) Junscheduled DNA synthesis assay, compound DE-79

**~28-0ay** 

### toxicity study in rats '13-Week feeding study in rats 'A range-finding teratology study in rats with DE-79



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# &-f:.;}. .October ll; 1976

.; ! :._1 Final Report
ltl Vl'rnO :.l!CROBIOI.OGICAL MUTAGENICITI(, STUDIES 

## 'OF 'ifcii:iit cuiA-GEI 

### Gi ;CORPORATION cm.IPOU.NDS .::


## • Prepared :for: 

# . f(.e

# '0-.br'dl~t/jc14.<~d~ 

# .·' cAr. 1:/1=. 

# 3'trt--sr- (, '**CIDA~GEIGY** CORPORATION ArdslP.y, New York 1.0502

A ttent19n: Dr. Martin G. Bernstein

Submitted by: · Vincent ·F. SJ.nunon, :.tnaager Microbid.l Genetics Program Denis 

## c. Poole M{c:robiologist
SRI Project LSC-5702
Approved: 

# ,,**if.!~** 

# w.**/frv~** Gordoh 

## ·w. Newell, Director Department of 'I'oxicolO'gy 

# vu. &

. w.- A. Skinner, Executive DircCto:i= 

### Life Sciences Division '() 

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### sni tes:tcd 

### :four CIBA-G:E:IGY**CorP,,or;~tion co::~poundn**  

### to evaluate ,their potential "for DUt:tceneSis in five 

### S"tr:11ns of 

### Salmonella 

### typhirnuriurn--TAl535 1 

### TA1537 1 

### TA1538 1 

### wu.-5 pe(t".formed in the presence and i.n 

### TA98 1 

### and TAlCO, Each**~ss~y** ' 0 .;:1 c. 

# the absence of**n~metnbolic** 

### system. Compound'42l-32-A was very .}' ' d) 

### mut:.genic "1n strains 

# ,,

nc:t1vat1on TA1'!;35 and TAlCO when 

### the**rat-liver~homogenate** 

### metabolic oct1**v~** 

### t.1an o 

# ' system**~fS** 

### added to the**aas~y.** 

### 7he three other compounds,**421-~2-B 1**  421-:32-c, 

### and !.!A-26 1 

### were 

### nssays 

# 1n**t~B** 

### presence on system.



## .. " .:: 

### not**mutage~c** 

### in any 

## o: the**bacteria~** " '' ' "-' " 

### " 

### in the absence ot a metabolic act1.yat1on**il2~-32B=(Hexabromocyc~ododecane** 

### No.**760403~8)** o


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,. IJ SRI examined :four CIBA-GEIGY Corpor.at:1.on compounds for r.nit::'.l'- gen1c1 ty by 

### .!,::**~** l':licro01.ologicOJ.l**as~.ays** w1 th 1'1.ve strains 

## ?:r S3lr.tonell:J typhimur1um. An Kroclor 1251-stimula

### t~·d~ rrit-liver

### homogenate metabolic activation**sy~tem** 

### was :1.ncluded in**~he** 

### assay .. 0



### procedure 

## • incapable to provide**metabo~ic** 

### steps that**the;bac~Cria** 

### e1ihe:r: are ·.f:: o:f condtict:1ng or do not 

### Carry out under the

## a~~3y cond.1 

### t!Ons.;.:- The P.urpose of this study was to determi.ne whether thtj**compound~** elicited a mutngenic response ;tn**miCroorganism~.** 

### The assay p;rfOcedure w1th S. typh!:muriurn has been proven t.o 

### be 85 

### to 90% 

### accurate 1n detecting carcinogens**:a~~** mutagens," and it b.:ls

.. • ? • ·- ·.about the saq1e accuracy in -1dentif;Yi.ng chemicOJ.ls that are- nOt Carci.no-·

# ~~nic .. - 1 

### How7ver, because the test

# \y~1:em .is not

# 100~ accura te 1 neither a 'pos1 ti ve nor a negative**re_~ponse** prC?ves th::1 t :l chem1.cal
c 

### 1s**h~zordous** 

### or nonh;Iz:.trdous to rnan.

2 

### ()**~-·** 

## .**··~ ·~:**  

### .. r.1ETitODS
Salmonelln typhimurlum Strfiins T•'l535, T,'\1537, TJ'l538,**TA!JS~** :md T;\100

..
The Salmonelln typhirn.urium st:ra1ns used at SRI :n·e nll histidine


### nu otrophs by

### virt~e**~:f** mutationS in the histid1"ee operon. When" these hiStidlne-depfmdent

### ce~ls**!.~:re** grown on 

# a rn1nimnl media petl·i plnte 

### deficient in h:1stid1ne', 

## • + independence (his ) are onl;r those cells that revert to hl.Stid

.. ine 0 '

. able to grow (mutants). The spontaneous mut.<l'"tion**!req~ency** 

## Or each strain 1s rel:ltively constant, but when


### a. mutagen is added**~0** the**:t{;':Jr~'the** mutation frequency is increased 

### 3 to ;J.OO times.

\'le obtained our!·**'~'typhirnul·ium** strnins from Dr

.. , Eruce Ames of the University of

### Cali:forn~a nt

### D~rkeley.**"::"s** In :Jddi_tion to having

### ~=tutations in the histidine operon, all the i.rid.icntor strains hnvc 

### mutations in the lipopolysacch::trlde coat (r.fa 

## -j :md deletions that cover a gene involved in the repair o! uv d:un:1ge**(~D-L** The :rfa- mutation makes •.i

-- the ntrains more perrneabtc. to l:xrg:e r:;olccu"les, thereby increasing their**~ensi** ti\.'1 

### ty to these molccti'lcs. 

## tJ The uvrB- mutation decreases repair o£ some types o! Chemically darn3ged

### D~A and thereby

### enh~nces**sens~'ti** Vi 

### ty to some**r.tut.;;~.genic** 

### chectic:llS·. Strairi TJ'l535 is 

### reverted



### . + ' to hiStidine prototrophy (his ) by foiany r.mtagcns t.h:Jt ,cause J>ase-pnir -:; ' ::J.: 

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## •

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.. substitutions.**7~100** 

## iS derived 

### from ·TA1535 by the introduction .:: the 

### n**·'fnc~or plas~~d**  ptu.tlOl. G The**i:~troduction** Of**~hiS** plasmid,

### ·-~~- 0 

### ·=**~** ··hi h 

### 'le c: a. ...

## ~.f t.)

## m·~~**,~ ~**  

# t**\1~** '.1 

### c ......**~** "!:.. 9:**~** c:: ;')

..... :'t' 

### c':'nfers ampicillin 

### r-esistance to the stt·ai:', 

### ¥':rcatly enhnncris tJ1e**~1**

### ~ens1tiv1.ty of the strnin to some

### b~se-~~i'~-.-~ubsti1:u1:ion**mu~::agens.. "~~·~8-~**  

## ..**~** \'le hz.ve shown 

### ihat mutagens such _as

### b~nzYi chlol?dc 

## omd 2-{2-furyi.)_- 3-(5-nitro-2-furyl) ocrylnmide (known'os AF2) con: be detected in plnte assays by TAlOO but not**b~·** 

### "f}\1535. The**pre:sence:.·~f** this 

### plasmip alsO makes strain T.\100 sensitive to ·same

## frame~hift**~~tngens--e.g·.,** 0 

### ICR-191,

. : 

### .. benzo(o)pyrene, alfatoxtn 

### EF and 7,12-dimethy1benz(a)nnthiitcene.**~** .:**~ '·~~~**  '**~** 

### a' 

## "**>:!·~~-** 

## · .

. \1 ;:**St:r~1ns** TJ\1537 ond 'TJ\1538 ::arc reve:rtod by**m~ny** frnmeshift mutogcns. T/

1 1537 is more

### sens~tive**th~n** TJ\1538 to muto:ltion by some acrldi.ncs and ben::anthracens. 7 but the difference is quantit:Jtive r.nther th::m qualita-

.. tive. TA9S is derived from T/\1.538 by the addition of the plasmid pi-::.MlOl, which makes this str.nin more sensitive to**~orne muta~;cns.**  

### .**~** All the indicator strains are stored ot 

### -80°Cn For each experiment, ::m 1i1oculum :from frozen stock cultures is grown ov:ern1ght at 37°C in a nutrient broth consisting of 1% 

### -tryptone :Jnd

### 0.5~ yeast extract. After stationary overnight growth, the cultures are shaken for 

### 3 to 

### 4 hours 

### to ensure optimal**gr~wtb.** Each 

### culture is checked for sensitivity to crystal violet. The presence of the rfa- mutation makes !i'the indicator strains sensi.tive to this dyr, whereas the parent 

### +
.. . strain, 

## !!! , is not sensitive to the dye.**Howeve~,** the mutation is 

### • ' + ' rcversi.ble, leading to the accumulation o:t: rfa cells i.n the culture. !) ·-- • ' Therefore,, the cells must be tested routinely to ensure thei-r sensitivity to c:r·yst:al v:1.olet. Eoch culture also is tested by specific mutagens known to revert each test strain ( pos1 ti.ve controls). AsSnys in Pgnr

To a sterile 13 

### x 100 l{tl'n test tube placed i.n a 43°C heating block., we add in the following order: 

### (l} (2) q 2 ml of 0.6% agar o 0

.. 05 ml o! indicator organ.isms (3) 0.5 ml of metabolic activation mixture (optional) (4) Up to 50 IJ.l of·' a solution of the test chemical.
For negative-controls, we use'steps 

## (1), (2}, and 

## (3) (optional) and 50 lJ.l o! the solvent Used for the test chemical.
This 

### mixture is sti.rred gently and then" poured onto r.tinimal agar

### plntes~t After 

### tt1e soft agar has set, the plates are incubated at 

# q. 6% agar contains 0.05 rn.\1 histidine and 0.05 m.\l biotin. 

### 1 Minimal agar plates consist of 15 :-g of agar, 20 g of glucose, 0.2 g 

## oi MgS0 4 '7 .H?O, 2 g o! citric ,acid monohyd:atc, 10 g of K 2 IIPo 4 , and**3.~** g of NafiNH 4 

### Po 4 ·H 2 o per liter.
4

' \, 

## \

.. 

## , b. 

### = '. 0

. ·=-

# .......... "

') 

### + The nur.1ber o! his tJ rcvert:an ts is coun·ted :md recorded

.. Some of the revert::mi.s ;:arc routinely tested to confirr.t that they arc 

### hts+, require b1ot1n 1 ::md nre .sensitive



### t~ cryst::tl violet (rf:a-) •

.. 

### , 

### . 

# --

**Mct:~bolic** 

### .ACti va tion--il

**rocl\J~"** 1254-Stimul:::J ted r.te;:nboli c

**~ct1** \."a t1on System Some carcinogenic 

### mutagens (e

.. g. 1 

### d1rnethyln1tros::am1ne) are 

### 1nnct1ve



### unlc:S~ they nre converted to their



### ~ct:i.ve fort:'i by betn'g metilbo11zed. 

### ftmes has described the

**rnet:::~bol1c** 

### nct1vat1on Systems we use.G1 7 Jldult male rats (250 to 300 

### g) are given a single 500 mg/kg intraperitoneal injcc'tion of a polychlorinated biphenyl (J'roclor 1254). 7 Four days_ 

### after the 1nj;ct1on, the 

### animals' :food 

### is z·emoved.. On the fifth cby, the 

### rats are 

### killed and 

### the 11ver homogenate prepared

.. The livers are removed asept1C4!lly and placed in prcwe1ghed sterile glass beakers. 'l'he org:n.n weight is 

### determined,

**:m~** 

### ::all subsequent operntions to the 

### metabolic activati?n step a:x·e conducted 1n an i:_-ce b3th.. The orr;.:Jn 1s w::1shed in ::10 equal volume of' cold, sterile 0

.. 15 M KCl (lml/g of wet or:;:m)., minced with sterile surbiCill scissors

.. in three volumes of 0.15

**K~l,** and

**ho~ogenized** with a **Potter-Elvehje~.** 

### apparatus. The

**hornogenat~·is centr1~uged**  fer 10 minutes

**~t** 9000

**x~**



### 1 and the supernatant 

### 1s removed and 

### stored 'in liquid nitrogen. 

### The met<lbolic acti\•ation system f'or e::ach experiment consists of (for 10 ml): 

### ('! 1) 1

.. 0 ml• postmitochrondrial sttpernat;J.n!= 2} 0.2 ml J.!gC1 2 (0.4 

### lot) l(Cl (1.65 

### :.t) 3} 0.05 

### ml t;lucose-6-phosphote ( 1 M) 4} 

### 5) 0.4 ml NP.DP. (0.1 

### lot) 5.0 ml 

### sodium phosphnte (0.2 M pH 7.4}



### 5 

## . '

### . 

# - .. : 

# ..

· Jtl-:SUJ:l'S .1\ND DISCUSSION"

ImmCdin tcly bciol"c '1scJ

**c~ch** compOund aiid the po::o:l tivc controls vterc <ll:.sol\·cd in dimethyi'sulioxidc



### (D~ISO) so that 50 Ill o! the solvent contnlncd the dcsirCct dose of



### chcr.~.1col. The negative control 

# ,, wns tl'C:l'l:cd wi lh 50 J.!.l of D.\150. fhc rcsul ts pres en ted in "fable 1 nrc the nvcr;:q;c of at le4lst two c_::tpcr.lmcn t:s conducted on scx1arn tc dnys. .An



### cxp~rimcn t consists of :.::;soJys with the lcs t compound nnd with posi t1 vc and n ega tl vc controls.

**Eac~** compound is tested first over a wlde rnf1gc of .,doses on nll str01ins in the presence ond in the absence of li vcr homoccnate for datcrmi.nation of its toxicity ;:md/or mutar:cnicity

.. li' the compound .c.Jocs not nppcar to be



### mut::~gcnic or



### tox~c, it is 

# " ·retested for confirmation of this result. Compounds 421-32-B_,. 421:--32-c, nnd MA-26 w&rc not mutagenic or to:x:ic.. However, the prcliminnry



### c:x:pC~imci1t wi.th compound -121-32-A indicated that it was



### mut::~cenic on st.r-.ains_ "''Al535 and TAlbO in the

**pr~sence** o:f the liver homor:cnate. Two additional experiments were pcr.formcd to confirm that



### 421-~2-A i.s mutnacnic.. Table l presents the nvcrage 

### oi these two experiments

.. Compound 421-32-A was verY mutag-enic when tested Wi.th ratliver-homogenate



### activa~i.on system, but it was not



### rnutngcn~c in the absence of ·the activation system. The 'compound was more mut<Jgcnlc in assays with TAlOO than with TA1.535.



### Fail~·ly linear dose responses were obtained in both Of these strains up to a dose of 100 l'g-/plate.. Compound

**~21-32-A** was wenl<ly mutor;enic in TA98 1 .tn "the presence o! the actiVation system.. St:rain T.,\1535 1.s 

### reverted· to histidine indcpenedence by mutagens that



### c~u;e base-pai.r substituti:on



### mut::~tions



## 1 whereas- strai.n TAlOO is 

### reverted 

## by bothfrnmeshift nnU base-pair substitution mutagens. 

### .-

6•.



# •

.· .... - r·•, ,; CC:uupc.•tl1at1

## ~::n:~J:.!-;\ J.::; n :;

## ll'OI~l!'**mut:~~c.n :t~ltl**  1::;: ·COIIl,ilill"OJIJlc J•clot'l.l vo lo m11lnscn1c nct:!vlty 

### :.md ns.:;ny cO'ndit:ionsi 

### (i.c. 1 

### it in 

### ruutact!nic

### 1~: ' -: :i I,' I • • s t.l>n1ns "li\1535 mal "J'J\.100 only when the li vor hoint)gcnil tc is

## pr<.:~cn t) 

### <r· to tris(2 1 

### 3 1 

### -<ll:bromopropyl) phospl1a ta 1

## ~nether haior;cnatcd

## al1ph~tc phoscphate we**h~·ve** 

### tested.



## '

{j ·)'



# •··

,c

:J



### .. 6 

## a."''};,'.,;:>":,-,.CompoUndlfecntivccontrolPostt1\•econtrol.;-PrOpiol:JctonoAF-29-JlD1noncr1d1no2-An,hram1naCln.I-GEIOY421-32-AMeto.bollcActivity++++++++++,•.,;..'...()'l'nblel!!!.AS&iYSWITU'riPHIMURIUM,,pr.ofCompoundliddedperPlato/lvcroge·NurnborofUistid.ine-PosttiveRevertantsporPlauTJ\1537TA153d.279282716519H-343716350712-7820,0510031283820·486320"5771380111101327170s102582.927·-1GSll,;::9":;;711N159101G271'10501005001000108ll331G5u8p"3·115221411251773G1110•2720211015272·15S27IS3G-J7810177111241.5'212541ol121135.8·1-1100sao10007!H0IS135-Il3.:l!l12151919702GOG233028199515002•JGO?/:.2922101<1200....•··"":'_,..··-',;......'..--....,•.'.•·.•'·'._,·>'"7-".;·-,,(r',·;·.:'.,.•'·'.•n(j"'•,•,,,,Table1(.cont1nuod)•'""'M(!tabolicJ!&olCompoundAvcror.cNU!nbcrofll1:Jtidine-Po!l1tivcncv(!rtont.sperPiatcCompound/JetlvityAddedperPlnto".ClDA-GtlGX421-32-D-140150::J529112..-.f:l10301527271535038·202227HD'.-10030132525163500371G30!GO10003G1G213915-1nooo·19101823157,,+13G214139l55+1034122530l-15+50,.;..31143G3G1<!9co+10028204139liJl,.31153740152+10003G123·1+50001313212G1G3421-32-C-143122937l;!:J10291429271375030IG2G311631003G1<12831l-17sOb32101<1321-1110004012213G1'11sooo..2372123181..+140154G1G130+103G103G4t1-1!)+5030103939l3G+1003018433G151+'50033IS393Gl::!G,-:_:;_+1000431838•1313G+5000.IG152330130,.•.. ) compound

Cl ll/.•OEI OY MA-26

"' A :3'

..... -:• C") 0"

. g: r.f· ;r a--

- ::0 3 I!J

**~ ~**  **o·~**

"''!.. .u.. !.?.,_g .- .. iJ Mctoba11c .. **-~:** Aetivlty ·- +

+

+

+

+

+

+ " **-~** Q **~:.:** . Tnb1o 1 (Concludedi •

1-lC of Compound

J\dded per Plnto •: _.

l

10

so

100

500

. 1000 5000 1

10 --

**c~** 50

100

500

1000

5000

" - Avcroce Number of lltst1dlne-Pos1ttve Revertants

**~ ~·r-**  **-~ ~** 

19 9 19 10

31 11 25 43

23 12 • 38 54

18 8 30 11

18 13 25 52

23 11 33 37

39 8 41 33

19 11 23 **'t~-** 36

11 a· 31

**3~**

21 15 39 14

14 **·.'~t4** 30 17

14 iii 36 11

16 18 31 15

19 ·n 12 15

" ,,

" "

" " J)cr Plato t.\-100 171

131

132

137

132

112

1G9;:;-:o<' !51

1:N

153

130

!57

!50

1<10 : h. 0 D .. i

" . . ,'! '• •

, .. ..

### ,.

• 

### ..


## m:n:m·ll cES

., 1. J. McC:-mn, E. Cho1 1 E. Ynr:msak1 1 and B. N. Ames. Detection of 

### carcinoccns as

**mut~~cns** 

### in the Salmonclln microsome 

### tCst: Assay of 300

**chcm1c~ls. Proc~.**  Jlnt. Acad. Sc1. USA

**~,** 5135-5139.

• ... 2. B. N

.. AmcsJ E. G

.. Gurney, J

.. A. Miller, and H. Dartsch. Carcinogens :: as frarncsh1:Ct mutagens: Mctabol1 tcs and d<!-riv3t1vcs of 2-acctylar.linoflttorcne and other aromatic amine cax·cinogcns. Proc. Nat. Acad. Sc1.



### USA~ 3128-3]32 (1972). " 3. B. N • .Ames" F. "D. Lc!!,

**an~** 

## w. E. Dur-ston. An improved

**b;:,cter1~1** 

### test system :Cor the detection and

**cl~ss~!1cat1on** 

### of

**rnuta~ens** 

### and carcinogens. Proc. Nat. Acad. Sc1. USA 70, 782-786 (1973). 

### 4. B. N. Ames, E. aJ·c mu tagcns: activation and

**~** (1973). :() 

### W. Durston, E. Yamasaki., 

### and F. D • .,Lee. Carcinogens A simple test system comb1n1ng liver homogenates for 

### bacteria for de'te:ct1on. Pro c. Nat

.. .Aoz::ad

.. Sci.. USA

5. J. McCann, N. E. Splng01:r., J

.. Kobox:.1 1 and B. N. Ames. The detection 

### o!

**corcino~ens** 

### as

**~Ut3ecns: Bacter1~l**  

### tester strains with F £actor p1asm1ds. P7oc.

**N~t.** Acod. Sc1. US' (1n press, 1975).

**6~** · L. D

... Kicr, E. Ya:r..o.saki 1 ::md B

... N. Ames. 

### activity in cigarette smoke condens3tes

.. 71, 4159-4163 (1974) •. De tcction of mutagenic. Proc. Nat. Acad. 5<:1., USA 

## ' · 7. -;B. N

.. Ames, 

## .;r. McCann., omd 

### E. Y01rnasak1.. l •. 1ethods 

### for Detecting Carcinogens. and Mutagens wl th the

**Salr.~onella/M::~li'l.mal1:ln-M1crosome** Mutngen1c1ty Test. J,lut. Res. 31 7 347-364 (1975) 0 ••



# ...

. :;



### 10

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## ·'

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.... LLI



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